CN109423493B - Cold-resistant gene OSRYH1 of rice and application thereof - Google Patents

Cold-resistant gene OSRYH1 of rice and application thereof Download PDF

Info

Publication number
CN109423493B
CN109423493B CN201710787875.4A CN201710787875A CN109423493B CN 109423493 B CN109423493 B CN 109423493B CN 201710787875 A CN201710787875 A CN 201710787875A CN 109423493 B CN109423493 B CN 109423493B
Authority
CN
China
Prior art keywords
rice
cold
osryh1
gene
resistant
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CN201710787875.4A
Other languages
Chinese (zh)
Other versions
CN109423493A (en
Inventor
李成刚
刘勇
王丹
张德咏
谭新球
王国梁
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
HUNAN PLANT PROTECTION INSTITUTE
Hunan Agricultural University
Original Assignee
HUNAN PLANT PROTECTION INSTITUTE
Hunan Agricultural University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by HUNAN PLANT PROTECTION INSTITUTE, Hunan Agricultural University filed Critical HUNAN PLANT PROTECTION INSTITUTE
Priority to CN201710787875.4A priority Critical patent/CN109423493B/en
Publication of CN109423493A publication Critical patent/CN109423493A/en
Application granted granted Critical
Publication of CN109423493B publication Critical patent/CN109423493B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8273—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for drought, cold, salt resistance

Landscapes

  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • Wood Science & Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Botany (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Plant Pathology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Microbiology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicinal Chemistry (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention discloses a cold-resistant gene of rice and application thereof, wherein the nucleotide sequence of the cold-resistant gene OSRYH1 of rice is a sequence shown as SEQ ID NO.1, and the amino acid sequence of the encoded protein of the cold-resistant gene is a sequence shown as SEQ ID NO. 2. The rice cold-resistant gene OSRYH1 and ubi promoter drive the OSRYH1 gene, so that the over-expressed transgenic plant has obvious cold-resistant effect and can be applied to improving the cold-resistant performance of plants. Meanwhile, OSRYH1 is derived from rice, so that the rice transformed with OSRYH1 gene has reduced concerns about food safety.

Description

Cold-resistant gene OSRYH1 of rice and application thereof
Technical Field
The invention relates to the technical field of genes, in particular to a cold-resistant gene OSRYH1 of rice and application thereof.
Background
The rice is the staple food of more than half of the population all over the world, but the global climate constantly changes, and extreme weather often happens, and low-temperature cold damage can occur every year to rice production, seriously influences the output, endangers the grain safety. According to statistics, the rice yield loss caused by low-temperature cold damage in China is about 10% every year. Research shows that the cold resistance of rice is controlled by gene, so that the development of cold resistance gene and the improvement of the cold resistance of rice are effective ways of coping with low-temperature cold damage and ensuring the safety of rice production and grain.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a cold-resistant gene OSRYH1 of rice and application thereof, the cold-resistant gene OSRYH1 of rice can obviously improve the cold resistance of rice, the survival rate of the rice with the gene OSRYH1 is more than 90% after the rice is treated at low temperature for 3 days and recovered at normal temperature for 7 days, and the gene OSRYH1 of rice can be applied to preparing transgenic cold-resistant rice.
In order to solve the technical problems, the invention provides a cold-resistant gene OSRYH1 of rice, wherein the nucleotide sequence of the cold-resistant gene OSRYH1 of rice is a sequence shown as SEQ ID NO. 1; the amino acid sequence of the coding protein of the cold-resistant gene OSRYH1 of the rice is the sequence shown in SEQ ID NO. 2.
As a general technical concept, the invention also provides an application of the cold-resistant gene OSRYH1 of rice in improving the cold resistance of rice.
The application method preferably comprises the following steps:
s1, designing a primer according to the cold-resistant gene OSRYH1 of the rice, and carrying out PCR amplification to clone OSRYH 1;
s2, constructing the clone OSRYH1 into a vector pCAMBIAubi1300 to obtain a pCAMBIAubi1300-OSRYH1 vector;
s3, transforming the pCAMBIAubi1300-OSRYH1 vector into Nipponbare rice by utilizing an agrobacterium transgenic method to obtain cold-resistant rice.
In the above application method, preferably, in the primers, the upstream primer is a sequence shown by SEQ ID No.3, and the downstream primer is a sequence shown by SEQ ID No. 4.
Compared with the prior art, the invention has the advantages that:
(1) the invention provides a cold-resistant gene OSRYH1 of rice, a ubi promoter drives the gene OSRYH1, the cold-resistant effect of an over-expressed transgenic plant is obvious, and the gene can obviously improve the cold resistance of the rice in southern double cropping rice cultivation areas which are easily subjected to cold damage in China and rice planting areas which are easily subjected to abnormal weather, and can respond to sudden cold damage in the growth period of the rice and ensure the production of the rice. The cold resistance of the rice transformed with the OSRYH1 gene is obviously improved, the survival rate is more than 90 percent after the rice is recovered at normal temperature for 7 days after being processed at low temperature for 3 days, and the survival rate of the contrast is less than 5 percent (figure 1). The gene is applied to genetic improvement of rice, and can greatly improve the cold resistance of the rice.
(2) The invention provides a cold-resistant gene OSRYH1 of rice, which is derived from rice, so that worry about food safety of the rice transformed with the gene OSRYH1 is reduced. The deep research on the gene can reveal the molecular mechanism of regulating and controlling the cold resistance of rice and provide new gene resources for the genetic improvement of the cold resistance of rice and other plants.
Drawings
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention.
FIG. 1 is a graph showing the expression level of OSRYH1 along with the time of cold stress in example 1 of the present invention.
FIG. 2 is a graph showing the results of the expression levels of LOC _ Os03g09140 in roots, stems and leaves in example 1 of the present invention.
FIG. 3 is a graph showing the results of cold treatment of cold-resistant rice in example 2 of the present invention.
Detailed Description
The invention is further described below with reference to the drawings and specific preferred embodiments of the description, without thereby limiting the scope of protection of the invention.
The materials and equipment used in the following examples are commercially available.
Example 1:
an identification of a cold-resistant candidate gene OSRYH1 of rice.
The cold-resistant gene OSRYH1 of rice in this example is located at the cold-resistant QTL site of rice located on the third chromosome by whole genome association analysis, and further the difference between the gene sequences of the cold-resistant population and the cold-sensitive population is analyzed and found by
LOC _ Os03G09140 lacks 27bp in a cold-sensitive variety, is positioned in the third intron region of the gene, is therefore determined as a candidate gene, belongs to ras small G protein gene, has high homology with ryh1 gene in yeast and is named as OSRYH 1. Then, it was further confirmed from the results of experiment 1 and experiment 2 that OSRYH1 is a cold-resistant candidate gene.
The OSRYH1 gene sequence is a nucleotide sequence shown as SEQ ID NO.1, and specifically comprises the following steps:
ATGGCGCCGGTGGTGTCGGCGCTGGCCAAGTACAAGCTGGTGTTCCTCGGCGACCAGGCGGTGGGGAAGACGGCCATCATCACCCGCTTCATGTACGACAAGTTCGACGCCACCTACCAGGCGACGATCGGGATCGATTTCCTCTCCAAGACGATGTACCTCGAAGACCGAACCGTACGGCTGCAGCTTTGGGATACAGCTGGGCAGGAGAGGTTCCGGAGCTTGATCCCAAGTTACATCAGGGATTCTTCTGTGGCAGTGATTGTTTACGATGTGACTGACAGGCAGTCCTTTTTGAGTATATCGAAGTGGATTGAAGAGGTAAATACACAACGAGGCGGAGATGTGATCATCGTCCTTGTTGGAAATAAAACAGACCTTGTTGACAAGAGGCAAATATCCACTGACGAAGGAGAAGCCAAGGCACAGGAGCACGGCGCCATGTTCATGGAAACCAGCGCAAAAGCTGGATTCAACATCAAGCCACTGTTTCGCAAGATCGCTGCATCGCTGCCTGGAATGGAGGCGCTCTCATCGGCAAAGCAGGAAGACATGGTGGACATAAACCTACGGCCTGCTGCCTCCGGGCAAATACCTTCAGGGGCAGAAGCACAAGAAGAACAGAAAGCAGGCGGTTGTTCCTGCTGA。
experiment 1: analyzing the expression trend of the cold-resistant gene OSRYH1 of the rice.
(1) Putting 3-week-old Nipponbare rice seedlings into a light incubator for cold treatment at 8 ℃, 12H light and 75% humidity for 3 days, and taking leaf samples at 0 hour, 12 hours, 24 hours, 36 hours, 48 hours and 72 hours respectively.
(2) Total RNA in leaf samples was extracted separately and reverse transcribed to synthesize the first strand cDNA.
(3) Primers were synthesized based on the cDNA sequence of the LOC _ Os03g09140 gene, and the expression of LOC _ Os03g09140 at each treatment period was detected using Real-time PCR.
See figure 1 for results: the expression level of OSRYH1 was greatly increased with the increase of cold stress time, indicating that OSRYH1 was induced by cold stress.
Experiment 2: specific expression of the rice cold-resistant gene OSRYH1 in root, stem and leaf tissues is analyzed.
(1) Taking the roots, stems and leaves of the Japanese fine rice which is 3 weeks old.
(2) Extracting total RNA from root, stem and leaf samples respectively, and performing reverse transcription to synthesize a first strand of cDNA.
(3) The specificity of expression of LOC _ Os03g09140 in root, stem and leaf tissues under normal growth conditions was also examined.
See figure 2 for results: LOC _ Os03g09140 is shown to be expressed in significantly higher amounts in roots than in stems and leaves.
Example 2:
an application of a cold-resistant gene OSRYH1 of rice in preparing cold-resistant rice.
On the basis of example 1, in this example, OSRYH1 is used to construct an overexpression vector, and then a transgenic technology is used to prepare cold-resistant rice. The method of the embodiment comprises the following steps:
(1) and (3) extracting total RNA of leaves of the Japanese fine rice.
1.1, taking 0.1g of Nipponbare rice leaves, grinding the leaves in liquid nitrogen, and transferring the leaves into a 1.5mL RNAase-free centrifuge tube. Adding 1.0mL of Trizol extracting solution into an RNAase-free centrifuge tube, fully whirling, shaking and mixing uniformly to obtain a mixed solution I.
1.2, continuously adding 200 mu L of chloroform into the mixed solution I, and fully vortexing and oscillating to obtain a mixed solution II.
1.3, centrifuging the mixed solution II at 12000rpm for 10min at 4 ℃. And adding the centrifuged supernatant into another 1.5mL RNAase-free centrifuge tube, adding isopropanol with the volume equivalent to 2/3 of the supernatant, and uniformly mixing to obtain a mixed solution III.
1.4, the mixed solution is placed on ice for 30min, and centrifuged at 12000rpm for 10min at 4 ℃. After centrifugation, the supernatant was removed, and the precipitate was rinsed by adding 1mL of 70% ethanol, centrifuged at 12000rpm for 5 minutes, and the supernatant was removed. The centrifuged precipitate was left at room temperature for 5 minutes to remove 70% ethanol, and 50. mu.L of RNase-free ddH was added2And O, dissolving the precipitated total RNA.
The content of the extracted total RNA was measured at 342 ng/. mu.L and stored in a freezer at-80 ℃.
(2) First strand cDNA Synthesis.
2.1, the cDNA synthesis reaction system is as follows:
2X qPCR reverse transcription Mix: 10 mu L of the solution;
total RNA (50-1000 ng): 2 mu L of the solution;
H2O：8μL。
2.2, reaction procedure: the first strand cDNA is obtained by storing at 25 deg.C for 10min, 50 deg.C for 30min, and 85 deg.C for 5min at 4 deg.C. After first strand cDNA synthesis, the cDNA was stored at-20 ℃ in a freezer.
(3) Cloning of the OSRYH1 Gene: the OSRYH1 gene was cloned using RT-PCR technique. The method specifically comprises the following steps:
3.1, primer design:
forward primer (Forward primer): 5'-GGATCCATGGCGCCGGTGGTGTC-3' (SEQ ID NO. 3);
reverse primer (downstream primer): 5'-AAGCTTTCAGCAGGAACAACCGCCTG-3' (SEQ ID NO. 4).
3.2, RT-PCR reaction system:
2×KOD buffer：25μL；
10mM dNTPs：1μL：
10mM Forward primer：1μL：
10mM Reverse primer：1μL：
KOD DNA polymerase：1μL：
cDNA template (first strand cDNA obtained in step (2)): 1 mu L of the mixture is added into the solution,
ddH2O：20μL。
3.3, reaction program: pre-denaturation at 95 ℃ for 3 min; denaturation at 95 ℃ for 30 s; annealing at 55 deg.C for 30 min; extension is carried out for 4s at 68 ℃, and the steps of cyclic denaturation, annealing and extension are carried out for 35 times; final extension at 68 ℃ for 5 min; and storing at 4 ℃ to obtain an RT-PCR product.
And 3.4, carrying out electrophoresis and detection on the RT-PCR product in 1% agarose gel, cutting a target band, and recovering by using a gel recovery kit to obtain a recovered product.
(4) And (5) constructing a transformation vector.
4.1, cloning the recovered band into pEASY-Blunt vector by using T-zero kit to obtain reaction product of the recovered product in step (4).
Reaction system: PCR product 4.5. mu.L (40-50ng), pEASY-Blunt 0.5. mu.L.
4.2, mixing the reaction products obtained in the step 5.1, centrifuging for 5s for a short time, and standing for 15min at 25 ℃.
4.3 and then transforming JM109 Escherichia coli competent cells with a heat shock method to obtain a transformant.
4.4 transformants were cultured at 37 ℃ for 1h, then spread evenly on 50mg/L kanamycin (kanamycin) LB plates, and cultured at 37 ℃ for 16 h.
And 4.5, selecting a single clone, and carrying out colony PCR detection to obtain a positive clone.
Performing DNA sequencing on the cloned OSRYH1 gene, wherein the DNA sequence of the OSRYH1 gene is shown as SEQ ID NO: 1, specifically:
ATGGCGCCGGTGGTGTCGGCGCTGGCCAAGTACAAGCTGGTGTTCCTCGGCGACCAGGCGGTGGGGAAGACGGCCATCATCACCCGCTTCATGTACGACAAGTTCGACGCCACCTACCAGGCGACGATCGGGATCGATTTCCTCTCCAAGACGATGTACCTCGAAGACCGAACCGTACGGCTGCAGCTTTGGGATACAGCTGGGCAGGAGAGGTTCCGGAGCTTGATCCCAAGTTACATCAGGGATTCTTCTGTGGCAGTGATTGTTTACGATGTGACTGACAGGCAGTCCTTTTTGAGTATATCGAAGTGGATTGAAGAGGTAAATACACAACGAGGCGGAGATGTGATCATCGTCCTTGTTGGAAATAAAACAGACCTTGTTGACAAGAGGCAAATATCCACTGACGAAGGAGAAGCCAAGGCACAGGAGCACGGCGCCATGTTCATGGAAACCAGCGCAAAAGCTGGATTCAACATCAAGCCACTGTTTCGCAAGATCGCTGCATCGCTGCCTGGAATGGAGGCGCTCTCATCGGCAAAGCAGGAAGACATGGTGGACATAAACCTACGGCCTGCTGCCTCCGGGCAAATACCTTCAGGGGCAGAAGCACAAGAAGAACAGAAAGCAGGCGGTTGTTCCTGCTGA。
the coding amino acid sequence of OSRYH1 gene is shown as SEQ ID NO: 2, specifically:
MAPVVSALAKYKLVFLGDQAVGKTAIITRFMYDKFDATYQATIGIDFLSKTMYLEDRTVRLQLWDTAGQERFRSLIPSYIRDSSVAVIVYDVTDRQSFLSISKWIEEVNTQRGGDVIIVLVGNKTDLVDKRQISTDEGEAKAQEHGAMFMETSAKAGFNIKPLFRKIAASLPGMEALSSAKQEDMVDINLRPAASGQIPSGAEAQEEQKAGGCSC。
4.6, selecting positive clones, extracting plasmid DNA, carrying out double enzyme digestion on BamH1 and HindIII, and recovering the gene fragment OSRYH 1. The pCAMBIAubi1300 vector was double digested with BamH1 and HindIII and recovered. The OSRYH1 gene fragment was ligated to the same double-digested pCAMBIAubi1300 vector to obtain pCAMBIAubi1300-OSRYH1 vector.
Connecting a reaction system: OSRYH 1: 2 mu L of the solution; pcambiubi 1300: 2 mu L of the solution; t4DNA ligase: 0.5 mu L; t4DNA ligase buffer: 0.5. mu.L.
And (3) connecting: 22 ℃ for 3 h.
4.7 the pCAMBIAubi1300-OSRYH1 vector was transformed into competent cells of Escherichia coli JM109 by heat shock method, and then applied evenly to 50mg/L kanamycin (kanamycin) LB plate and cultured at 37 ℃ for 16 hours. Then selecting a single clone, extracting plasmid DNA in the single clone, carrying out double enzyme digestion detection by using BamH1 and HindIII, and selecting a positive clone for the transgenosis of rice.
(5) Obtaining the transgenic rice plant with the OSRYH1 gene.
5.1, introducing the pCAMBIAubi1300-OSRYH1 vector into an Agrobacterium EHA105 strain, infecting mature embryo callus of the Nipponbare rice, and obtaining resistant callus through co-culture, recovery culture and screening culture (the hygromycin concentration in the screening culture medium is 50 mg/L).
And 5.2, culturing the resistant callus through differentiation, rooting and the like to obtain a regeneration plant.
And 5.3, transplanting the regenerated plants into a nutrition pot, and planting the plants in a greenhouse. Taking leaves of the survived regenerated plants, extracting DNA, and detecting the regenerated plants by using a PCR technology, wherein the plants containing the OSRYH1 gene are positive trans-OSRYH 1 gene plants.
Experiment 3: and verifying the cold resistance of the transgenic plant.
Positive transgenic OSRYH1 gene plants were planted in nutrition pots, 6 plants were planted in each pot, and the repetition was repeated 3 times, and normal growth was carried out in a greenhouse for 3 weeks using japanese fine rice as a control. Then, 3 weeks of the size of the plants were subjected to 8 ℃ cold treatment. The cold treatment is carried out for 3d, the normal temperature is recovered for 7d, and the phenotype is observed.
See figure 3 for results: the leaves of the positive transgenic OSRYH1 plant were found to be still green, whereas the leaves of the control Nipponbare rice plants had withered. The result shows that the cold resistance of the rice can be obviously improved by the over-expression of the OSRYH1 gene. After 3 times of repeated experiments, the OsRYH1 gene-transformed rice shows strong cold resistance. Therefore, the experimental result proves that the OSRYH1 gene can obviously improve the cold resistance of rice and is a new gene for improving the cold resistance of rice.
The foregoing is merely a preferred embodiment of the invention and is not intended to limit the invention in any manner. Although the present invention has been described with reference to the preferred embodiments, it is not intended to be limited thereto. Those skilled in the art can make many possible variations and modifications to the disclosed embodiments, or equivalent modifications, without departing from the spirit and scope of the invention, using the methods and techniques disclosed above. Therefore, any simple modification, equivalent replacement, equivalent change and modification made to the above embodiments according to the technical essence of the present invention are still within the scope of the protection of the technical solution of the present invention.
Sequence listing
<110> institute of plant protection in Hunan province, university of Hunan agriculture
<120> cold-resistant gene OSRYH1 of rice and application thereof
<130> do not
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 648
<212> DNA
<213> Nippon fine Rice (Rice)
<220>
<221> misc_feature
<222> (1)..(648)
<223> OSRYH1 Gene sequence
<400> 1
atggcgccgg tggtgtcggc gctggccaag tacaagctgg tgttcctcgg cgaccaggcg 60
gtggggaaga cggccatcat cacccgcttc atgtacgaca agttcgacgc cacctaccag 120
gcgacgatcg ggatcgattt cctctccaag acgatgtacc tcgaagaccg aaccgtacgg 180
ctgcagcttt gggatacagc tgggcaggag aggttccgga gcttgatccc aagttacatc 240
agggattctt ctgtggcagt gattgtttac gatgtgactg acaggcagtc ctttttgagt 300
atatcgaagt ggattgaaga ggtaaataca caacgaggcg gagatgtgat catcgtcctt 360
gttggaaata aaacagacct tgttgacaag aggcaaatat ccactgacga aggagaagcc 420
aaggcacagg agcacggcgc catgttcatg gaaaccagcg caaaagctgg attcaacatc 480
aagccactgt ttcgcaagat cgctgcatcg ctgcctggaa tggaggcgct ctcatcggca 540
aagcaggaag acatggtgga cataaaccta cggcctgctg cctccgggca aataccttca 600
ggggcagaag cacaagaaga acagaaagca ggcggttgtt cctgctga 648
<210> 2
<211> 215
<212> PRT
<213> Nippon fine Rice (Rice)
<220>
<221> SITE
<222> (1)..(215)
<223> amino acid sequence encoding OSRYH1 Gene
<400> 2
Met Ala Pro Val Val Ser Ala Leu Ala Lys Tyr Lys Leu Val Phe Leu
1 5 10 15
Gly Asp Gln Ala Val Gly Lys Thr Ala Ile Ile Thr Arg Phe Met Tyr
20 25 30
Asp Lys Phe Asp Ala Thr Tyr Gln Ala Thr Ile Gly Ile Asp Phe Leu
35 40 45
Ser Lys Thr Met Tyr Leu Glu Asp Arg Thr Val Arg Leu Gln Leu Trp
50 55 60
Asp Thr Ala Gly Gln Glu Arg Phe Arg Ser Leu Ile Pro Ser Tyr Ile
65 70 75 80
Arg Asp Ser Ser Val Ala Val Ile Val Tyr Asp Val Thr Asp Arg Gln
85 90 95
Ser Phe Leu Ser Ile Ser Lys Trp Ile Glu Glu Val Asn Thr Gln Arg
100 105 110
Gly Gly Asp Val Ile Ile Val Leu Val Gly Asn Lys Thr Asp Leu Val
115 120 125
Asp Lys Arg Gln Ile Ser Thr Asp Glu Gly Glu Ala Lys Ala Gln Glu
130 135 140
His Gly Ala Met Phe Met Glu Thr Ser Ala Lys Ala Gly Phe Asn Ile
145 150 155 160
Lys Pro Leu Phe Arg Lys Ile Ala Ala Ser Leu Pro Gly Met Glu Ala
165 170 175
Leu Ser Ser Ala Lys Gln Glu Asp Met Val Asp Ile Asn Leu Arg Pro
180 185 190
Ala Ala Ser Gly Gln Ile Pro Ser Gly Ala Glu Ala Gln Glu Glu Gln
195 200 205
Lys Ala Gly Gly Cys Ser Cys
210 215
<210> 3
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<220>
<221> misc_feature
<222> (1)..(23)
<223> designed according to the experimental requirements, as the upstream primer for PCR.
<400> 3
ggatccatgg cgccggtggt gtc 23
<210> 4
<211> 26
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<220>
<221> misc_feature
<222> (1)..(26)
<223> designed according to the experimental requirements, as a downstream primer for PCR.
<400> 4
aagctttcag caggaacaac cgcctg 26

Claims (1)

1. An application of a cold-resistant gene OSRYH1 of rice in improving the cold resistance of rice, wherein the nucleotide sequence of the cold-resistant gene OSRYH1 of rice is a sequence shown as SEQ ID NO. 1; the amino acid sequence of the coding protein of the cold-resistant gene OSRYH1 of the rice is the sequence shown in SEQ ID NO. 2.
CN201710787875.4A 2017-09-04 2017-09-04 Cold-resistant gene OSRYH1 of rice and application thereof Expired - Fee Related CN109423493B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710787875.4A CN109423493B (en) 2017-09-04 2017-09-04 Cold-resistant gene OSRYH1 of rice and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710787875.4A CN109423493B (en) 2017-09-04 2017-09-04 Cold-resistant gene OSRYH1 of rice and application thereof

Publications (2)

Publication Number Publication Date
CN109423493A CN109423493A (en) 2019-03-05
CN109423493B true CN109423493B (en) 2021-08-13

Family

ID=65513819

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710787875.4A Expired - Fee Related CN109423493B (en) 2017-09-04 2017-09-04 Cold-resistant gene OSRYH1 of rice and application thereof

Country Status (1)

Country Link
CN (1) CN109423493B (en)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110373424B (en) * 2019-07-09 2022-12-23 西藏自治区农牧科学院农业研究所 Production method of cold-resistant rice containing HVUL5H49059.2 gene
CN112321691A (en) * 2020-11-10 2021-02-05 广东省农业科学院水稻研究所 Application of OsGF14f protein in regulation and control of cold resistance of rice
CN113637685B (en) * 2021-08-26 2023-04-18 湖南农业大学 Cold-resistant gene OsRab11C1 of rice and application thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101037696A (en) * 2006-03-16 2007-09-19 华中农业大学 Paddy cool injury gene and application

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101037696A (en) * 2006-03-16 2007-09-19 华中农业大学 Paddy cool injury gene and application

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Genome-wide Association Mapping of Cold Tolerance Genes at the Seedling Stage in Rice;Dan Wang等;《Rice》;20161115;第9卷;第1-10页 *
登录号：XM_015772737;佚名;《GenBank》;20160301;第239-886位 *

Also Published As

Publication number Publication date
CN109423493A (en) 2019-03-05

Similar Documents

Publication Publication Date Title
CN107541520B (en) OsSAUR11 gene related to rice root development and stress resistance, coding protein and application
CN109423493B (en) Cold-resistant gene OSRYH1 of rice and application thereof
CN113265408B (en) Pseudo-ginseng DOF transcription factor gene PnDof1 and application thereof
CN112501182A (en) Poplar ERF transcription factor gene and application thereof
CN115873086A (en) Tomato transcription factor SlWOX13 gene and protein and application thereof
CN109879947B (en) Phyllostachys pubescens transcription factor PheDof2 gene and application thereof
CN108715852B (en) Tomato fruit mature gene Sl0658 and application thereof
CN107663524B (en) FvGAIP gene for regulating and controlling strawberry stolon generation and application thereof
CN110004173B (en) Method for obtaining non-transgenic storable fresh-eating Chinese wolfberry
CN114214332B (en) Tianmu rehmannia anthocyanin related gene RcMYB1 and application thereof
CN106434659B (en) Soybean low-temperature inducible promoter, recombinant expression vector containing promoter and application
CN114807165A (en) Application of corn ZmNAC78 gene
CN108103075B (en) Switchgrass gene PvC3H29 for delaying plant senescence and application thereof
CN106701780B (en) PgAG gene for regulating development of pomegranate ovule and application thereof
CN118324889B (en) PheARF1.5 protein related to high regulation and control of phyllostachys pubescens stalk, coding gene and application
CN112458103B (en) Gene for regulating and controlling capsorubin accumulationCaBBX20And uses thereof
CN114015666B (en) Application of OsPARP3 gene in regulation and control of plant drought tolerance
CN114875043B (en) Betula alba BpPIF4 gene participating in adventitious root development and application thereof
CN113549602B (en) Phyllostachys pubescens ascorbic acid peroxidase gene PeAPX1 and application thereof
CN112979777B (en) Application of OsWRKY74 gene of rice in regulation and control of seed germination and pre-harvest sprouting
CN114395022B (en) Apple zinc finger protein transcription factor MdZF-HD11 and application thereof
CN113278631B (en) Pear PbPRR2 and PbPRR3 genes and application thereof
CN113046364B (en) Application of rice OsATL9 gene in regulation of rice resistance
CN113046365B (en) Application of rice OsATL17 gene in regulation of rice resistance
CN117187261A (en) Pear ERF transcription factor PpERF098 and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20210813